Jiangsu Province Key Laboratory of Anesthesiology, Jiangsu Province Key Laboratory of Anesthesia and Analgesia Application Technology and NMPA Key Laboratory for Research and Evaluation of Narcotic and Psychotropic Drugs, Xuzhou Medical University.
Jiangsu Province Key Laboratory of Anesthesiology, Jiangsu Province Key Laboratory of Anesthesia and Analgesia Application Technology and NMPA Key Laboratory for Research and Evaluation of Narcotic and Psychotropic Drugs, Xuzhou Medical University;
J Vis Exp. 2021 Apr 19(170). doi: 10.3791/62254.
Alzheimer's disease (AD) is a neurodegenerative disease that contributes to 60-70% dementia around the world. One of the hallmarks of AD undoubtedly lies on accumulation of amyloid-β (Aβ) in the brain. Aβ is produced from the proteolytic cleavage of the beta-amyloid precursor protein (APP) by β-secretase and γ-secretase. In pathological circumstances, the increased β-cleavage of APP leads to overproduction of Aβ, which aggregates into Aβ plaques. Since Aβ plaques are a characteristic of AD pathology, detecting the amount of Aβ is very important in AD research. In this protocol, we introduce the immunofluorescent staining method to visualize Aβ deposition. The mouse model used in our experiments is 5×FAD, which carries five mutations found in human familial AD. The neuropathological and behavioral deficits of 5xFAD mice are well-documented, which makes it a good animal model to study Aβ pathology. We will introduce the procedure including transcardial perfusion, cryosectioning, immunofluorescent staining and quantification to detect Aβ accumulation in 5×FAD mice. With this protocol, researchers can investigate Aβ pathology in an AD mouse model.
阿尔茨海默病（AD）是一种神经退行性疾病，占全球痴呆症的 60-70%。AD 的一个显著特征无疑在于脑内淀粉样蛋白-β（Aβ）的积累。Aβ是由β-淀粉样前体蛋白（APP）经β-分泌酶和γ-分泌酶的蛋白水解切割产生的。在病理情况下，APP 的β-切割增加导致 Aβ的过度产生，Aβ聚集形成 Aβ斑块。由于 Aβ斑块是 AD 病理的特征之一，因此检测 Aβ的含量在 AD 研究中非常重要。在本方案中，我们介绍了免疫荧光染色法来可视化 Aβ的沉积。我们实验中使用的小鼠模型是 5×FAD，它携带了在人类家族性 AD 中发现的五种突变。5xFAD 小鼠的神经病理学和行为缺陷已有详细记录，使其成为研究 Aβ病理学的良好动物模型。我们将介绍包括心脏灌注、冷冻切片、免疫荧光染色和定量检测 5×FAD 小鼠 Aβ积累的程序。通过该方案，研究人员可以在 AD 小鼠模型中研究 Aβ病理学。
